GSM6767339: Patient 19, Primary tumor; Homo sapiens; RNA-Seq
After disruption and homogenization of the fresh frozen tumor samples, RNA was extracted using the Qiagen RNeasy Plus Mini kit. Paraffin embedded tissue block (FFPE) blocks were sectioned to prepare unstained sections (5μm thick) on glass slides for RNA extraction. For FFPE samples, after scraping the tumor tissue present on 15 slides with a razor blade, samples were de-paraffinized using the Qiagen Deparaffinization Solution and RNA was extracted using the Qiagen RNeasy FFPE kit. RNA yield and purity were measured photometrically with NanoDrop and Qubit Fluorometer. RNA quality was analyzed using the Fragment Analyzer System. Total RNA libraries were prepared using Illumina stranded Total RNA prep ligation with ribo-zero plus kit starting with an input amount of 200 ng total RNA. Library size and quality were confirmed on Fragment Analyzer System. KAPA quantification kit for Illumina platforms was used to quantify libraries. Samples were pooled in equimolar fashion. Ten pb double indexed libraries were sequenced on a NextSeq 550 instrument using 74 base-length read chemistry in paired-end mode.